Department of Gynecologic Oncology, Guangxi Medical University Cancer Hospital, Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor, Ministry of Education, Nanning, Guangxi, China.
J Ovarian Res. 2021 May 15;14(1):67. doi: 10.1186/s13048-021-00807-y.
LAMA3 is a widely studied methylated gene in multiple tumors, but the relationship between chemotherapy resistance in ovarian cancer is unclear. In this study, LAMA3 methylation was predicted by bioinformatics, and the ability of LAMA3 methylation to predict the chemotherapy resistance and prognosis of ovarian cancer was confirmed in experiments.
Multiple databases have performed the bioinformatics analysis of methylation and transcription factor binding site (TFBS) on the promoter region of LAMA3 gene. Pyrosequencing detected the methylation of LAMA3. QRT-PCR and immunohistochemistry detected the expression of LAMA3. Real Time Cell Analyzer (RTCA) detects changes in cell proliferation, migration and invasion ability. Flow cytometry was used to detect apoptosis.
CPG islands of 176 bp, 134 bp, 125 bp and 531 bp were predicted in the promoter region of LAMA3 gene. The 4 prediction results are basically overlapped. 7 transcription factor binding sites were predicted, and the one with the highest score was on the predicted CpG island located in the proximal promoter region. LAMA3 hypermethylation and low expression are both associated with chemotherapy resistance and poor prognosis in ovarian cancer. LAMA3 methylation was negatively correlated with expression. After upregulation of LAMA3, the proliferation ability of chemoresistant ovarian cancer cell decreased, while the ability of apoptosis, invasion and migration increased.
LAMA3 hypermethylation is associated with chemotherapy resistance and poor prognosis. As a typical CpG island gene, LAMA3(cg20937934) and LAMA3(cg13270625) hypermethylation is negatively correlated with low expression. LAMA3 promotes the invasion, migration and apoptosis of SKOV3DDP. In the future, the mechanism of LAMA3 methylation in ovarian cancer will need to be further studied.
LAMA3 是多种肿瘤中广泛研究的甲基化基因，但与卵巢癌化疗耐药的关系尚不清楚。本研究通过生物信息学预测 LAMA3 甲基化，并通过实验证实 LAMA3 甲基化预测卵巢癌化疗耐药和预后的能力。
多个数据库对 LAMA3 基因启动子区的甲基化和转录因子结合位点（TFBS）进行了生物信息学分析。焦磷酸测序检测 LAMA3 的甲基化。实时荧光定量 PCR（QRT-PCR）和免疫组织化学检测 LAMA3 的表达。实时细胞分析（RTCA）检测细胞增殖、迁移和侵袭能力的变化。流式细胞术检测细胞凋亡。
在 LAMA3 基因启动子区预测到 176bp、134bp、125bp 和 531bp 的 CpG 岛。4 个预测结果基本重叠。预测到 7 个转录因子结合位点，得分最高的一个位于位于近端启动子区的预测 CpG 岛上。LAMA3 高甲基化和低表达均与卵巢癌的化疗耐药和不良预后相关。LAMA3 甲基化与表达呈负相关。上调 LAMA3 后，耐药卵巢癌细胞的增殖能力下降，而凋亡、侵袭和迁移能力增加。
LAMA3 高甲基化与化疗耐药和不良预后相关。作为一个典型的 CpG 岛基因，LAMA3（cg20937934）和 LAMA3（cg13270625）的高甲基化与低表达呈负相关。LAMA3 促进 SKOV3DDP 的侵袭、迁移和凋亡。未来需要进一步研究 LAMA3 甲基化在卵巢癌中的作用机制。
